Munc18-1 induces conformational changes of syntaxin-1 in multiple intermediates for SNARE assembly

Abstract In neuronal exocytosis, SNARE assembly into a stable four-helix bundle drives membrane fusion. Previous studies have revealed that the SM protein Munc18-1 plays a critical role for precise SNARE assembly with the help of Munc13-1, but the underlying mechanism remains unclear. Here, we used...

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Autores principales:	Sanghwa Lee, Jonghyeok Shin, Younghun Jung, Heyjin Son, Jaeil Shin, Cherlhyun Jeong, Dae-Hyuk Kweon, Yeon-Kyun Shin
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9b8cf37ff71545a589930604bb4e66e8
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spelling	oai:doaj.org-article:9b8cf37ff71545a589930604bb4e66e82021-12-02T15:32:59ZMunc18-1 induces conformational changes of syntaxin-1 in multiple intermediates for SNARE assembly10.1038/s41598-020-68476-32045-2322https://doaj.org/article/9b8cf37ff71545a589930604bb4e66e82020-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-68476-3https://doaj.org/toc/2045-2322Abstract In neuronal exocytosis, SNARE assembly into a stable four-helix bundle drives membrane fusion. Previous studies have revealed that the SM protein Munc18-1 plays a critical role for precise SNARE assembly with the help of Munc13-1, but the underlying mechanism remains unclear. Here, we used single-molecule FRET assays with a nanodisc membrane reconstitution system to investigate the conformational dynamics of SNARE/Munc18-1 complexes in multiple intermediate steps towards the SNARE complex. We found that single Munc18-1 proteins induce the closed conformation of syntaxin-1 not only in the free syntaxin-1 but also in the t-SNARE (syntaxin-1/SNAP-25) complex. These results implicate that Munc18-1 may act as a gatekeeper for both binary and ternary SNARE complex formation by locking the syntaxin-1 in a cleft of Munc18-1. Furthermore, the kinetic analysis of the opening/closing transition reveals that the closed syntaxin-1 in the syntaxin-1/SNAP-25/Munc18-1 complex is less stable than that in the closed syntaxin-1/Munc18-1 complex, which is manifested by the infrequent closing transition, indicating that the conformational equilibrium of the ternary complex is biased toward the open conformation of syntaxin-1 compared with the binary complex.Sanghwa LeeJonghyeok ShinYounghun JungHeyjin SonJaeil ShinCherlhyun JeongDae-Hyuk KweonYeon-Kyun ShinNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-8 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sanghwa Lee Jonghyeok Shin Younghun Jung Heyjin Son Jaeil Shin Cherlhyun Jeong Dae-Hyuk Kweon Yeon-Kyun Shin Munc18-1 induces conformational changes of syntaxin-1 in multiple intermediates for SNARE assembly
description	Abstract In neuronal exocytosis, SNARE assembly into a stable four-helix bundle drives membrane fusion. Previous studies have revealed that the SM protein Munc18-1 plays a critical role for precise SNARE assembly with the help of Munc13-1, but the underlying mechanism remains unclear. Here, we used single-molecule FRET assays with a nanodisc membrane reconstitution system to investigate the conformational dynamics of SNARE/Munc18-1 complexes in multiple intermediate steps towards the SNARE complex. We found that single Munc18-1 proteins induce the closed conformation of syntaxin-1 not only in the free syntaxin-1 but also in the t-SNARE (syntaxin-1/SNAP-25) complex. These results implicate that Munc18-1 may act as a gatekeeper for both binary and ternary SNARE complex formation by locking the syntaxin-1 in a cleft of Munc18-1. Furthermore, the kinetic analysis of the opening/closing transition reveals that the closed syntaxin-1 in the syntaxin-1/SNAP-25/Munc18-1 complex is less stable than that in the closed syntaxin-1/Munc18-1 complex, which is manifested by the infrequent closing transition, indicating that the conformational equilibrium of the ternary complex is biased toward the open conformation of syntaxin-1 compared with the binary complex.
format	article
author	Sanghwa Lee Jonghyeok Shin Younghun Jung Heyjin Son Jaeil Shin Cherlhyun Jeong Dae-Hyuk Kweon Yeon-Kyun Shin
author_facet	Sanghwa Lee Jonghyeok Shin Younghun Jung Heyjin Son Jaeil Shin Cherlhyun Jeong Dae-Hyuk Kweon Yeon-Kyun Shin
author_sort	Sanghwa Lee
title	Munc18-1 induces conformational changes of syntaxin-1 in multiple intermediates for SNARE assembly
title_short	Munc18-1 induces conformational changes of syntaxin-1 in multiple intermediates for SNARE assembly
title_full	Munc18-1 induces conformational changes of syntaxin-1 in multiple intermediates for SNARE assembly
title_fullStr	Munc18-1 induces conformational changes of syntaxin-1 in multiple intermediates for SNARE assembly
title_full_unstemmed	Munc18-1 induces conformational changes of syntaxin-1 in multiple intermediates for SNARE assembly
title_sort	munc18-1 induces conformational changes of syntaxin-1 in multiple intermediates for snare assembly
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/9b8cf37ff71545a589930604bb4e66e8
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Munc18-1 induces conformational changes of syntaxin-1 in multiple intermediates for SNARE assembly

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